Antibodies for modulating binding between lrp and wise

Abstract

The present invention provides, inter alia, antibodies that modulate binding between Lrp5 and WISE or Lrp6 and WISE, but do not modulate binding between Lrp4 and WISE. Also provided are pharmaceutical compositions and kits containing such antibodies. Further provided are methods for preventing WISE binding to Lrp5 or Lrp6, but not WISE binding to Lrp4.

What is claimed is: 
     
         1 . A method for producing a monoclonal antibody, comprising the steps of:
 (a) injecting a laboratory animal with an effective amount of an antigen comprising one or more sequences selected from the group consisting of SEQ ID NOs: 1-48;   (b) isolating B cells from the animal's spleen;   (c) fusing the B cells isolated in step (b) with immortalized cells;   (d) incubating fused cells in step (c) in a medium to select hybridoma cells;   (e) screening for hybridoma cells that produce an antibody that binds to the selected one or more sequences from the group consisting of SEQ ID NOs: 1-48; and   (f) growing the chosen hydridoma cells to obtain the desired monoclonal antibody.   
     
     
         2 . The method of  claim 1 , wherein the laboratory animal is a mouse. 
     
     
         3 . The method of  claim 1 , wherein the antigen comprises one or more sequences selected from the group consisting of SEQ ID NOs: 1-39. 
     
     
         4 . The method of  claim 1 , wherein the antigen comprises one or more sequences selected from the group consisting of SEQ ID NOs: 1-15. 
     
     
         5 . The method of  claim 1 , wherein the antigen comprises one or more sequences selected from the group consisting of SEQ ID NOs: 1-3 and 16-27. 
     
     
         6 . The method of  claim 1 , wherein the antigen comprises one or more sequences selected from the group consisting of SEQ ID NOs: 1-3 and 28-39. 
     
     
         7 . The method of  claim 1 , wherein the antigen comprises one or more sequences selected from the group consisting of SEQ ID NOs: 40-48. 
     
     
         8 . A monoclonal antibody produced by a method according to  claim 1 . 
     
     
         9 . A method of detecting an antibody that binds to Lrp5 and/or Lrp6 but not Lrp4, comprising the steps of:
 (a) contacting antibodies with a pool of short peptides derived from Lrp4, Lrp5 and Lrp6;   (b) identifying antibodies that (i) specifically binds to Lrp5 or Lrp6 at one or more sequences selected from the group consisting of SEQ ID NOs: 1-3, 16-41, 43-44 and 46-47, and (ii) does not bind to Lrp4-derived peptides; and   (c) from the antibodies identified in step (b), determining and selecting the antibody that has specific binding activity (K a ) of at least 10 9  mol −1 .